| _version_ | 1866901702564642816 |
|---|---|
| author | Prasad, Akanksha |
| author_facet | Prasad, Akanksha |
| contents | <p>Biologics manufacturing is rapidly evolving to meet global</p> <p>demand for mRNA therapeutics, recombinant proteins,</p> <p>monoclonal antibodies, and vaccines. However, current</p> <p>bottlenecks—including long cycle times, costly reagents, and</p> <p>inconsistent yields—limit scalability and responsiveness.</p> <p>Addressing these challenges requires an integrated strategy</p> <p>spanning upstream synthesis, capping efficiency, purification, and</p> <p>scale-up.</p> <p>For mRNA production, innovations in <strong>template design </strong>such as</p> <p>plasmids with built-in poly(A) tails and minicircle DNA reduce</p> <p>enzymatic steps and impurities, streamlining in vitro transcription</p> <p>(IVT). Process variability can be further minimized by</p> <p>applying <strong>design-of-experiments </strong>approaches and <strong>real-time PAT</strong></p> <p><strong>tools </strong>(Raman, inline UV), enabling adaptive IVT control and</p> <p>improving reproducibility. <strong>CleanCap® technology </strong>offers a one-</p> <p>pot solution for co-transcriptional capping, achieving 90–99%</p> <p>efficiency and mimicking natural Cap1 structures. While specific</p> <p>variants such as CleanCap M6 have shown lower efficiencies</p> <p>under certain conditions, optimization and comparability studies</p> <p>support its advancement as a superior alternative to enzymatic</p> <p>capping.</p> <p>Protein and vaccine platforms benefit from <strong>advanced process</strong></p> <p><strong>controls </strong>for nutrient and oxygen regulation, engineered cell lines</p> <p>for consistent glycosylation, and perfusion culture systems that</p> <p>deliver three- to four-fold higher productivity compared to fed-</p> <p>batch.</p> <p>Downstream, <strong>integrated purification strategies</strong>—including</p> <p>mixed-mode chromatography for dsRNA removal, RNase-assisted</p> <p>pre-treatment, tangential flow filtration, and continuous</p> <p>chromatography—improve yield, reduce impurities, and cut</p> <p>operational timelines. Process intensification approaches such as</p> <p>inline buffer conditioning and closed transfer systems further</p> <p>reduce facility footprint and costs.</p> <p>Finally, <strong>digital enablers </strong>including AI/ML models, digital twins,</p> <p>and modular single-use systems are redefining scale-up and</p> <p>technology transfer by predicting risks, ensuring regulatory</p> <p>compliance, and improving global supply resilience.</p> |
| format | Recurso digital |
| id | zenodo_https___doi_org_10_5281_zenodo_19063529 |
| institution | Zenodo |
| language | |
| publishDate | 2025 |
| publisher | Zenodo |
| record_format | zenodo |
| spellingShingle | Enabling Efficient Drug Substance Manufacturing: mRNA and Biologics – From Reaction to Purification Prasad, Akanksha <p>Biologics manufacturing is rapidly evolving to meet global</p> <p>demand for mRNA therapeutics, recombinant proteins,</p> <p>monoclonal antibodies, and vaccines. However, current</p> <p>bottlenecks—including long cycle times, costly reagents, and</p> <p>inconsistent yields—limit scalability and responsiveness.</p> <p>Addressing these challenges requires an integrated strategy</p> <p>spanning upstream synthesis, capping efficiency, purification, and</p> <p>scale-up.</p> <p>For mRNA production, innovations in <strong>template design </strong>such as</p> <p>plasmids with built-in poly(A) tails and minicircle DNA reduce</p> <p>enzymatic steps and impurities, streamlining in vitro transcription</p> <p>(IVT). Process variability can be further minimized by</p> <p>applying <strong>design-of-experiments </strong>approaches and <strong>real-time PAT</strong></p> <p><strong>tools </strong>(Raman, inline UV), enabling adaptive IVT control and</p> <p>improving reproducibility. <strong>CleanCap® technology </strong>offers a one-</p> <p>pot solution for co-transcriptional capping, achieving 90–99%</p> <p>efficiency and mimicking natural Cap1 structures. While specific</p> <p>variants such as CleanCap M6 have shown lower efficiencies</p> <p>under certain conditions, optimization and comparability studies</p> <p>support its advancement as a superior alternative to enzymatic</p> <p>capping.</p> <p>Protein and vaccine platforms benefit from <strong>advanced process</strong></p> <p><strong>controls </strong>for nutrient and oxygen regulation, engineered cell lines</p> <p>for consistent glycosylation, and perfusion culture systems that</p> <p>deliver three- to four-fold higher productivity compared to fed-</p> <p>batch.</p> <p>Downstream, <strong>integrated purification strategies</strong>—including</p> <p>mixed-mode chromatography for dsRNA removal, RNase-assisted</p> <p>pre-treatment, tangential flow filtration, and continuous</p> <p>chromatography—improve yield, reduce impurities, and cut</p> <p>operational timelines. Process intensification approaches such as</p> <p>inline buffer conditioning and closed transfer systems further</p> <p>reduce facility footprint and costs.</p> <p>Finally, <strong>digital enablers </strong>including AI/ML models, digital twins,</p> <p>and modular single-use systems are redefining scale-up and</p> <p>technology transfer by predicting risks, ensuring regulatory</p> <p>compliance, and improving global supply resilience.</p> |
| title | Enabling Efficient Drug Substance Manufacturing: mRNA and Biologics – From Reaction to Purification |
| url | https://doi.org/10.5281/zenodo.19063529 |